We will develop optimized methods for the enzymatic production of modified oligonucleotides which have strong advantages as therapeutic agents and biosensors. We will first implement a Sequential Injection Analysis (SIA) platform with continuous data collection to optimize biochemical protocols with large parameter spaces. We will use this platform to optimize conditions for in vitro transcription of 2'-O-methyl and 2'-fluoro RNA. We will also implement a continuous, continuous- or semi-continuous-flow bioreactor for in vitro production of modified RNA for use in ongoing projects including microfluidic cancer cell capture and in vivo evaluations of aptamer therapeutics. Microreactors of the type we will build have been demonstrated for controlled, efficient, scalable, synthetic organic chemistry. These same advantages make the strategy attractive for biochemical reactions. PUBLIC HEALTH RELEVANCE: Chemically modified DNA and RNA are more resistant to being destroyed by the environment in blood, and so they are preferred as tools to develop drugs or instruments used for clinical testing. Current methods for generating these valuable substances are very expensive and produce very little of the product, which limits their use. This project will develop an improved method to generate modified DNA and RNA which will allow wider use of these important chemical tools.